I. Body fluids - Must remain constant - Intake = output Body Fluid Dynamics Metabolic H 2 0 Preformed Free H 2 0 in Urine Feces Sweat H 2 0 Food - Body fluid distribution: Insensible H 2 0 loss Capillary Membrane 1/3 ECF Plasma 3L Interstitial Fluid 11L Lymphatic Circulation Cell Membrane 2/3 ICF Intracellular H 2 0 28L 70 Kg Person 42 Kg H 2 0 (60% BW) ~ 42L - Edema = excess fluid in body tissues o Intracellular o Extracellular Causes: Increase in Capillary in starling forces Filtration Coef. Lymphatic - cap. pressure - Histamine system - oncotic pressure - Bradykinin blockage - Capillary Perm.
- Fluid composition a. Intracellular b. Extracellular i. [K+] Plasma Intersitial ii. [Protein] Protein Protein iii. [Na+], [Cl-], [Ca++] cations - Measuring fluid volumes - Indicator dilution principle Inject 10 mg of indicator Volume x concentration Mass = Vol. x conc. (indic) therefore 10mg = vol. x (.01mg/L) - assumptions: 1. indicator has to spread throughout entire compartment 2. indicator has to disperse only in compartment of interest 3. Indicator is not metabolized or excreted [Indicator] - Plasma Volume use radio labeled albumin - ECF use substance that will not spread to cells (sucrose, radio labeled Na+ or Cl- - Total body water use tritium or deuterium oxide - ICF by subtraction II. Kidneys - Regulation of long term blood pressure - Regulation of RBC production (EPO) - Regulation of H20 electrolyte balance - Regulation of osmolarity - Regulation of acid/base balance - Excretion of metabolic waste and foreign chemicals - Glucose synthesis - Ca++ and P regulation - Vitamin D production A. Nephron Time
- functional unit of the kidney (composed of glomerulus and tubule) - ~ 1 million/kidney - not regenerated - 2 major components: 1. Glomerulus Renal corpuscle - Glomerular capillary bed - Bowman s capsule 2. Tubule Distal Convoluted Tubule Glomerular Capillaries Bowman s Capsule Proximal Convoluted Tubule Collecting Duct Loop of Henle: - descending limb - ascending limb (thin) - ascending limb (thick) - Types of nephrons: 1. Cortical = outer cortex; short loops of henle 2. Juxtamedullary = long loops of henle - help to concentrate urine - vasa recta = special capillary bed running in same direction as loop of henle B. Glomerular Capillary Membrane Fenestrations in capillary endothelium Collagen Lumen of Bowman s Capsule Podocytes Filtration Slits
III. Renal Physiology - ~1200mL of blood can move past glomeruli each minute - ~650mL is plasma - ~125 ml of the plasma is forced into renal tubules (equivalent to filtering the entire plasma volume up to 60x/day). - b/c of this much activity, the kidney s use ~20-25% of all O 2 in the body. - Filtrate formed is the precursor to urine - filtrate = everything in plasma (minus proteins) by the time it reaches the collecting ducts almost all H20, nutrients, and ions have been removed. - after all molecules are removed, this is considered urine. - Kidneys filter ~180L/day of this amount, only 1% usually leaves the body as urine the other 99% returns to the general circulation. - There are 3 major processes leading to urine formation: 3 options here: o Glomerular filtration - filtered; not reabsorbed o Tubular reabsorption - filtered; reabsorbed o Tubular secretion - filtered; secreted (K+) A. Glomerular Filtration - Passive process in which hydrostatic pressure forces fluids and solutes through the membrane. - Why is the glomerulus so efficient? o Membrane has S.A. and is highly permeable to H20 and solutes. o Glomerular BP is much higher than in other capillary beds; results in net filtration pressure. - Since plasma proteins are too large to move out of capillaries this maintains the colloid osmotic pressure (Π cap ) of glomerular blood. - Maintaining the Π cap prevents the loss of H20 into the renal tubules o P GC = Blood pressure in glomerular capillaries ~ 55mmHg o Π GC = Osmotic pressure in glomerular capillaries ~ 32mmHg - If plasma proteins are in the urine, this is usually indicative of a malfunction in the filtration membrane. - So what is Net Filtration Pressure (NFP)? o Pressure responsible for filtrate formation o Involves the glomerular hydrostatic pressure (HP G or P GC ) Chief factor pushing H20/solutes out of blood and across the filtration membrane. o However, theoretically, the colloid osmotic pressure in Bowman s capsule (Π BC ) should pull the filtrate into the
tubule this pressure is essentially 0mmHg b/c no proteins enter the capsule. o HP G is opposed by 2 factors: Colloid osmotic pressure of glomerular blood (OP G /Π GC ) Capsular hydrostatic pressure (HP c or P BC ) o Using the value for each, NFP can be calculated: NFP = P GC (Π GC + P BC ) NFP = 55 mmhg (30 mmhg + 15 mmhg) NFP = 10 mmhg!!! - So how much filtrate is formed each minute? o This is the glomerular filtration rate, or GFR. o 3 factors influence GFR: S.A. available for filtration Membrane permeability * NFP * This can be represented by the formula: - GFR = K f x NFP - Where K f is the capillary filtration coefficient, a measure of memb. Permeability. o So Long term regulation of GFR K f or in filtration pressure B.P. P GC GFR 10mmHg P GC ΠGC P BC NFP = 10mmHg P GC = 60mmHg P BC = 18mmHg Π GC = 32mmHg Π BC = 0mmHg
- s in resistance of arterioles also influences GFR: GFR (ml/min) Aff. Art. Eff. Art. Resistance - another impt. Feature of GFR is that it can be maintained over a range of blood pressures: Renal Blood Flow GFR Blood Pressure Renal Blood Flow - An B.P. = GFR - However, autoregulation of this process helps maintain GFR levels - There are 2 mechanisms to maintain GFR (intrinsic): o Myogenic B.P. Aff. Art. Pressure NFP GFR Stretch of Sm. Muscles Ca 2+ influx Aff. arterial constriction 2 Cell Populations: o Tubuloglomerular Feedback Juxtaglomerular apparatus: 1. JG cells = Pressure 2. MD cells = [Na + ]; flow rate Juxtaglom. Cells Macula Densa Cells Filtration Pressure DCT
Stimuli = B.P. or Tubular Flow Rate Glomerular Pressure (P GC ) GFR MD Cells - Dilate aff. Art. JG Cells: - activate RAA system - Angio. II const. eff. Art. - There are also 2 extrinsic controls: o SANS (stress response) Major effect = constrict arterioles Causes an in reabsorption and filtration Epi/NE bind to α-ar in sm. muscle o Hormonal Control RAA system (Angiotensin II) Overall = Na + and H 2 O reabsorption o Stimulates aldosterone prod/rel. o Constricts eff. Art. o Directly Na + reab. in proximal tubule by activating Na-K pump o Stimulates release of ADH Aldosterone Steroid hormone rel. from adrenal cortex Acts on principle cells in dist. tubule/cort. collecting duct. Na + reabsorption and K + secretion o Stimulates Na + -K + -ATPase on basolateral membrane o Na + channels or carrier proteins on luminal membrane ADH Peptide hormone rel. from posterior pituitary H 2 O permeability in distil tubule o Insert aquaporins in luminal memb.
Stimuli = B.P. or [Na + ] Rel. RAA Produces Angiotensin II BP, Na, H 2 O Rel. ADH H 2 O Reab. Aldosterone Na + reab. ANP Stimuli = heart stretch Effects = Renin release Aldosterone release Dilate aff. art. GFR Urine output B. Tubular Reabsorption/Secretion - Both act to alter the concentration of the filtrate - ~180L/day filtered 178L/day reabsorbed - Tubular reabsorption is the movement of filtered solutes and H 2 0 from the tubule lumen into the plasma. Peritubular Capillary Lumen of tubule Epithelial Cells IF Tubule Lumen Apical Surface Transcellular (solutes) Paracellular (H 2 O)
- Mechanisms: o Diffusion down gradients e.g. H 2 0 by osmosis o Active transport Primary e.g. Na + o Basolateral side Na-K pump o Luminal side Facilitated diffusion Secondary e.g. Glucose/Amino Acids o Na-K pump provides stored energy o Glu-Na cotransport/symport on luminal side o Glu moves down [ ] gradient into blood A.A. Na + A.A. K + Na + Na + Na + Na + G Na + G - Transport (tubular) maximum = the maximum amt. of a subst. that can be transported across the tubule membrane per unit time. - Sometimes there is excess glucose that cannot be reabsorbed due to renal threshold. - Renal Threshold = tubular load (mg/min) that exceeds the transport max. 3-2- - Other examples include: PO 4 and SO 4
C. Nephron Anatomy - Proximal Tubule o Largest reabsorbing segment (65-70% of filtrate) o Most secretion occurs here o Characteristics: Brush border on luminal membrane S.A. for reab. density of carrier proteins mitochondria Na-K ATPase Leaky epithelial. Junctions o Na +, H 2 0, Glu, Vitamins - Descending Thin Limb o Few mitochondria o No brush border o Active Transport o Permeable to H 2 0 + urea - Ascending Thick Limb diluting segment o Impermeable to H 2 0 + urea o Active Transport Na-K-2Cl cotransporter (reabsorption) Na-H counter transport (Na reabs; H secreted) Reabsorption of Ca 2+ and Mg 2+ - Distal Tubule site for regulation o Early DT = diluting Impermeable to H 2 0 + urea Site of JG apparatus (macula densa cells) Active Transport o Late DT/Connecting Tubule/Cortical Collecting Duct Impermeable to urea Permeability to H 2 0 varies w/o ADH impermeable w/adh permeable Principle Cells Na + reabsorption Both are acted on by K + secretion aldosterone Intercalated Cells K +, HCO 3 reabsorption H + secretion - Medullary Collecting Duct o Last segment to act on filtrate o Permeable to H 2 0, controlled by ADH o Permeable to urea (allows for reabsorption) o H + secretion
Proximal Tubule Distil Tubule Connecting Tubule Capillaries Glomerulus Bowman s Capsule Afferent/Efferent Arterioles Cortical Collecting Duct Cortex Medulla Descending Limb Medullary Collecting Duct Ascending Thick Limb D. Regulation of Reabsorption - Local Factors: o Glomerulotubular balance GFR reabsorption o Peritubular Starling Forces e.g. Angiotensin II Reabsorption via constricting eff. Arterioles pressure in peritubular capillaries. - Nervous Control o See page 7 - Hormones (humoral factors) o See page 7 E. Clearance
- Volume of plasma that is cleared of a substance by the kidneys per unit time. - Units = ml/min o C x = U x (V x ) Px ; where: x = substance tested C = Clearance U = Urine concentration (mg/ml) V = Urine flow rate (ml/min) P = Plasma Concentration (mg/ml) - Any substance used to measure Clearance also measures GFR the substance can be filtered, but not reabsorbed or secreted (Inulin/Creatinine). o If reabsorption occurs: Clearance < GFR o If secreted into tubule: Clearance > GFR IV. Dilution of Urine - Due to absence of ADH; loss of excess H 2 0 through excretion in urine - Plasma osm = 300 mosm/l - Filtrate osm = 300 mosm/l ~100mOSM/L 300 Absence of ADH leads to dilute urine Alcohol inhibits the release of ADH H 2 0 H 2 0 Ions Ions
V. Concentration of Urine - Max OSM of urine in humans ~ 1200 mosm/l - Sea water ~ 2400 mosm/l o 1L = 2400 mosm of electrolytes o 2L of urine A. Requirements - ADH o To reabsorb H 2 0 - High osmolarity in renal medullary interstitial fluid Produced by countercurrent multiplier - CC flow in loop of Henle - Active Transport (Na-K-2Cl) - Diffusion of H 2 0; permeability - Urea contributes ~500 mosm/l to the Renal I.F. osmolarity B. Maintaining the Osmotic Gradient - Vasa Recta acts as the countercurrent exchanger (maintains, not creates) - At bottom of loop, Plasma osm = 1200 mosm/l - At top of ascending loop, Plasma osm = 310 mosm/l o Vasodilators [urine] - Other factors affecting the gradient: o Length of the loop of Henle
Longer = greater concentration of urine o % of nephrons that juxtamedullary (long loops) More = urine concentration o Diet Protein osmolarity of urine VI. Diuretics - Substance that increases urine volume output - Clinically used to edema and hypertension o Cheaper than Ca 2+ channel blockers or ACE inhibitors - Types: o Osmotic diuretics Agents that are filtered, but not readily reabsorbed e.g. manitol, sucrose, urea, *glucose diabetes o Loop diuretics Work in ascending thick limb of loop of Henle Block Na-K-2Cl cotransporter e.g. Lasix (furosemide) o Thiazides Inhibits Na-Cl reabsorption in early DT Clinical o Competitive inhibitors of Aldosterone e.g. Spironolactone (aldactone) act on principle cells to prevent reabsorption o Na- channel blockers Collecting ducts Prevents Na + from being reabsorbed o Carbonic Anhydrase Inhibitors Prevents reabsorption of Na + and HCO 3 Acidosis can occur o Alcohol ADH antagonist o Xanthenes Caffeine Theophylline VII. Control of Osmolarity of ECF - ~300 mosm/l; 142 mosm/l comes from Na + - Osmoreceptor-ADH System: o Main stimulus = in osmolarity o Receptor cells = Osmoreceptors; Neurons in hypothalamus OSM (ECF) Cells Shrink Nerve Impulse Supraoptic Nuclei H 2 0 reabsorbed Negative Feedback Posterior Pituitary release of ADH into blood H 2 0 Permeability Late DT and collecting duct
o Other stimuli for ADH release: Blood Pressure (from baroreceptors) Cardiac stretch (via low pressure receptors) - Thirst System: o Stimulus = in osmolarity of ECF o Receptors = Osmoreceptors OSM ECF Osmoreceptors will stimulate thirst; occurs via the preoptic thirst center in the hypothalamus o Secondary Stimuli Blood pressure; ECF volume Angiotensin II Dryness of mucus membranes